Passage and reversal effects on gene expression of bovine meniscal fibrochondrocytes.
Passage and reversal effects on gene expression of bovine meniscal fibrochondrocytes.
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DOI：
10.1186/ar2293
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发表时间：
2007
影响因子：
4.9
通讯作者：
Athanasiou KA
中科院分区：
文献类型：
--
作者：
Gunja NJ;Athanasiou KA
膝关节半月板含有混合细胞群，表现出成纤维细胞和软骨细胞的特征。半月板的组织工程研究和未来的治疗需要大量的细胞接种在支架上。为了实现这一点，单层扩增通常用作增加细胞数量的技术。然而，这些细胞的表型可能与原代群体的表型显著不同。本研究的目的是调查的变化，在椎间盘纤维软骨细胞的基因表达水平超过四个代次，使用定量实时逆转录聚合酶链反应。使用了来自牛中动脉内三分之二的细胞。研究了半月板中常见的四种细胞外基质（ECM）分子，即胶原I、胶原II、聚集蛋白聚糖和软骨寡聚基质蛋白（COMP）。此外，将原代和传代的半月板纤维软骨细胞置于涂有胶原蛋白I或聚集蛋白聚糖蛋白的表面上，以研究是否可以逆转传代引起的任何基因表达变化。胶原蛋白I的表达被发现增加的通道数，而胶原蛋白II和COMP的表达下降。胶原蛋白I和聚集蛋白聚糖表面涂层显示在传代细胞中分别下调和上调胶原蛋白I和COMP表达水平。然而，胶原蛋白II表达的减少不能被逆转的蛋白涂层。这些结果表明，尽管单层扩增导致椎间盘纤维软骨细胞中基因表达的显著变化，但蛋白涂层可用于恢复几种ECM分子的原代细胞表达。
The knee meniscus contains a mixed population of cells that exhibit fibroblastic as well as chondrocytic characteristics. Tissue engineering studies and future therapies for the meniscus require a large population of cells that are seeded on scaffolds. To achieve this, monolayer expansion is often used as a technique to increase cell number. However, the phenotype of these cells may be significantly different from that of the primary population. The objective of this study was to investigate changes in meniscal fibrochondrocytes at the gene expression level over four passages using quantitative real-time reverse transcriptase polymerase chain reaction. Cells from the inner two-thirds of bovine medial menisci were used. Four extracellular matrix (ECM) molecules, commonly found in the meniscus, were investigated, namely collagen I, collagen II, aggrecan and cartilage oligomeric matrix protein (COMP). In addition, primary and passaged meniscus fibrochondrocytes were placed on surfaces coated with collagen I or aggrecan protein to investigate whether any gene expression changes resulting from passage could be reversed. Collagen I expression was found to increase with the number of passages, whereas collagen II and COMP expression decreased. Collagen I and aggrecan surface coatings were shown to downregulate and upregulate collagen I and COMP expression levels, respectively, in passaged cells. However, decreases in collagen II expression could not be reversed by either protein coating. These results indicate that although monolayer expansion results in significant changes in gene expression in meniscal fibrochondrocytes, protein coatings may be used to regain the primary cell expression of several ECM molecules.
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影响因子：
2.8
作者：
Le Graverand, MPH;Reno, C;Hart, DA
通讯作者：
Hart, DA
影响因子：
2.8
作者：
Lee, DA;Noguchi, T;Bader, DL
通讯作者：
Bader, DL
影响因子：
2.9
作者：
CHEUNG, HS
通讯作者：
CHEUNG, HS
影响因子：
2.8
作者：
Kambic, HE;McDevitt, CA
通讯作者：
McDevitt, CA
影响因子：
64.5
作者：
BENYA, PD;SHAFFER, JD
通讯作者：
SHAFFER, JD